The diverse genetic alterations that drive non-small cell lung cancer (NSCLC) provide attractive targets for therapeutic intervention, and tyrosine kinase inhibitors targeting mutant EGFR and rearranged ALK have demonstrated considerable clinical efficacy. However, treatment acquired resistance is a recurring theme in TKI therapy, and furthermore, current targeted therapies are not effective in KRAS-mutant NSCLC. The central goal of this P01 program is to advance drug development for NSCLC by developing novel approaches to targeting the kinases that drive these cancers, as well as those that facilitate the epigenetic and transcriptional adaptation that promotes resistance to targeted therapies. The kinase targets we focus on include mutant EGFR, c-RAF, Mek1/2, and transcription-associated cyclin dependent kinases. Inhibitor discovery and optimization is much more efficient when grounded in direct biochemical assays with purified components and when illuminated by co-crystal structures of lead compounds. The role of the Structure and Biochemistry Core (Core B) is to support each of the three projects by providing purified recombinant kinases, expertise in inhibitor characterization, in vitro assays for compound binding, molecular modeling and virtual screening, and crystallographic structure determination of kinase inhibitor complexes. Centralizing these activities in the core will prevent duplication of effort and expertise in the individual projects and will leverage the resources and experience available in the laboratory of the core director. We have extensive experience in the structural biology and inhibition of kinases and long-standing collaborative ties with the project leaders and core directors.